4-methylsulfonyl-2-butenenitrile and its pharmaceutical use

ABSTRACT

The present invention is directed to a method of treating an inflammatory skin disease or disorder, such as dermatitis, psoriasis, acne, or rosacea. The method comprises administering to the subject 4-methylsulfonyl-2-butenenitrile, in an amount effective to reduce or eliminate the symptoms of the inflammatory skin disease or disorder. Topical administration and oral administration are preferred routes of administration.

This application is a divisional of U.S. application Ser. No.15/084,816, filed Mar. 30, 2016; which is a continuation-in-part ofPCT/US2014/058685, filed Oct. 1, 2014; which claims priority to U.S.Provisional Patent Application No. 61/887,776, filed Oct. 7, 2013. U.S.application Ser. No. 15/084,816 also claims the benefit of U.S.Provisional application No. 62/142,855, filed Apr. 3, 2015. Theabove-identified applications are incorporated herein by reference intheir entireties.

FIELD OF THE INVENTION

The present invention relates to 4-methylsulfonyl-2-butenenitrile and apharmaceutical composition comprising same. The present invention alsorelates to methods of using the compound for treating an inflammatoryskin disease or disorder, such as dermatitis, psoriasis, acne, orrosacea.

BACKGROUND OF THE INVENTION

Inflammation is a process by which microbes or tissue injury induce therelease of cytokines and chemokines from various cell types producingincreased blood vessel permeability, upregulation of endothelialreceptors, and thus increased egress of various cells of the innate andadaptive immune system which enter surrounding tissue and grosslyproduce the classical picture of inflammation, i.e. redness, swelling,heat and pain.

Inflammation is a localized reaction of live tissue due to an injury,which may be caused by various endogenous and exogenous factors. Theexogenous factors include physical, chemical, and biological factors.The endogenous factors include inflammatory mediators, antigens, andantibodies. Endogenous factors often develop under the influence of anexogenous damage. An inflammatory reaction is often followed by analtered structure and penetrability of the cellular membrane. Endogenousfactors, such as mediators and antigens define the nature and type of aninflammatory reaction, especially its course in the zone of injury. Inthe case where tissue damage is limited to the creation of mediators, anacute form of inflammation develops. If immunologic reactions are alsoinvolved in the process, through the interaction of antigens,antibodies, and autoantigens, a long-term inflammatory process willdevelop. Various exogenous agents, for example, infection, injury,radiation, also provide the course of inflammatory process on amolecular level by damaging cellular membranes which initiatebiochemical reactions.

Based on the physical causes, pain can be divided into three types:nociceptive, neuropathic, and mix-type.

Nociceptive pain is the term for pain that is detected by nociceptors.Nociceptors are free nerve endings that terminate just below the skin,in tendons, in joints, and in internal organs. Nociceptive paintypically responds well to treatment with opioids and NSAIDs. There areseveral types of nociceptive pain: somatic pain, visceral pain, andcutaneous pain. Visceral pain comes from the internal organs. Deepsomatic pain is initiated by stimulation of nociceptors in ligaments,tendons, bones, blood vessels, fasciae and muscles, and is dull, aching,poorly localized pain. Examples include sprains and broken bones.Superficial pain is initiated by activation of nociceptors in the skinor other superficial tissue, and is sharp, well-defined and clearlylocated. Examples of injuries that produce superficial somatic paininclude minor wounds and minor (first degree) burns. Nociceptive pain isusually short in duration and ends when the damage recovers. Examples ofnociceptive pain include postoperative pain, sprains, bone fractures,burns, bumps, bruises, and inflammatory nociceptive pain. Inflammatorynociceptive pain is associated with tissue damage and the resultinginflammatory process.

Neuropathic pain is produced by damage to the neurons in the peripheraland central nervous systems and involves sensitization of these systems.Because the underlying etiologies are usually irreversible, most ofneuropathic pain are chronic pain. Most people describe neuropathic painas shooting, burning, tingling, lancinating, electric shock qualities,numbness, and persistent allodynia. The nomenclature of neuropathic painis based on the site of initiating nervous system with the etiology; forexamples, central post-stroke pain, diabetes peripheral neuropathy,post-herpetic (or post-shingles) neuralgia, terminal cancer pain,phantom limb pain.

Mix-type pain is featured by the coexistence of both nociceptive andneuropathic pain. For example, muscle pain trigger central or peripheralneuron sensitization leading to chronic low back pain, migraine, andmyofacial pain.

Connective tissues are subjected to a constant barrage of stress andinjury. Acute or chronic impacts and the natural progression of variousdegenerative diseases all produce painful inflammation in joint regions,such as the neck, back, arms, hips, ankles and feet. These afflictionsare common and often debilitating.

There is a need for a composition and a method for treatinginflammation, inflammatory-related disorders, and pain. The compositionshould be economic and easy to manufacture, and the method should beeffective and have no significant side effects.

DETAILED DESCRIPTION OF THE INVENTION Definitions

“About” as used herein, refers to ±10% of the recited value.

“Pharmaceutically acceptable salts,” as used herein, are salts thatretain the desired biological activity of the parent compound and do notimpart undesired toxicological effects. Pharmaceutically acceptable saltforms include various crystalline polymorphs as well as the amorphousform of the different salts. The pharmaceutically acceptable salts canbe formed with metal or organic counterions and include, but are notlimited to, alkali metal salts such as sodium or potassium; alkalineearth metal salts such as magnesium or calcium; and ammonium ortetraalkylammonium salts, i.e., NX₄+(wherein X is C₁₋₄.

“Solvates,” as used herein, are addition complexes in which the compoundis combined with an acceptable co-solvent in some fixed proportion.

4-Methylsulfonyl-2-butenenitrile

The inventors have isolated and identified4-methylsulfonyl-2-butenenitrile. The compound or its pharmaceuticallyacceptable salt or solvate is effective for treating inflammation,inflammatory-related disorders, and pain.

4-Methylsulfonyl-2-butenenitrile has a formula weight of 145.18, and itstrans structure is shown below.

4-methylsulfonyl-2-butenenitrile

4-Methylsulfonyl-2-butenenitrile can be synthesized by the addition ofsodium methanesulfinate to 4-bromo-2-butenenitrile in an organic solventcontaining water such as aqueous ethanol. The requisite4-bromo-2-butenenitrile can be prepared by bromination of allyl cyanideto give 3,4-dibromobutanenitrile, followed by base-catalyzed eliminationof HBr to introduce a double bond between C2 and C3, which is expectedto be an approximately 1:1 (e.g., between 40:60 to 60:40) mixture of E-and Z-isomers.

The present invention is directed to a method for preparing4-methylsulfonyl-2-butenenitrile. The method comprises a step (a) ofreacting 4-bromo-2-butenenitrile with sodium methanesulfinate. Anysolvent or solvent mixture that is suitable for providing a homogeneoussolution of the reactants is suitable. For example, a water-miscibleorganic solvent such as ethanol, methanol, isopropanol, acetone,tetrahydrofuran, or dioxane, containing a small amount (1-10% or 1-5%)of water is a suitable solvent for the reaction. In one embodiment,ethanol:water in a ratio of 30-40:1 is used as a solvent for thereaction.

The method may further comprise a step (i), before the above step (a),of preparing the requisite 4-bromo-2-butenenitrile by first reactingbromine (Br₂) with allyl cyanide, then adding a basic solution. In thisstep, allyl cyanide is first brominated in organic solvents such aspetroleum ether and alcohol, and then HBr is eliminated from themolecule to form a double bond by introducing at least one equivalentof, preferably about one equivalent of, an appropriate base. In thisstep, an appropriate base that is compatible with the solvents in thereaction mixture is used. A preferred base is an ethanolic solution ofsodium ethoxide, which can be prepared by dissolving sodium metal inethanol. After isolation and purification of the product4-bromo-2-butenenitrile, it can be used in step (a).

Pharmaceutical Compositions

The present invention provides pharmaceutical compositions comprisingone or more pharmaceutically acceptable carriers and an active compoundof 4-methylsulfonyl-2-butenenitrile, or a pharmaceutically acceptablesalt or a solvate thereof. The pharmaceutical composition can includeone of the cis or trans isomers, or both isomers either equimolar, or ofdifferent amounts. The active compound or its pharmaceuticallyacceptable salt or solvate in the pharmaceutical compositions in generalis in an amount of about 0.01-20%, or 0.05-20%, or 0.1-20%, or 0.2-15%,or 0.5-10%, or 1-5% (w/w) for a topical formulation; about 0.1-5% for aninjectable formulation, 0.1-5% for a patch formulation, about 1-90% fora tablet formulation, and 1-100% for a capsule formulation.

In one embodiment, the active compound is incorporated into anyacceptable carrier, including creams, gels, lotions or other types ofsuspensions that can stabilize the active compound and deliver it to theaffected area by topical applications. In another embodiment, thepharmaceutical composition can be in a dosage form such as tablets,capsules, granules, fine granules, powders, syrups, suppositories,injectable solutions, patches, or the like. The above pharmaceuticalcomposition can be prepared by conventional methods.

Pharmaceutically acceptable carriers, which are inactive ingredients,can be selected by those skilled in the art using conventional criteria.Pharmaceutically acceptable carriers include, but are not limited to,non-aqueous based solutions, suspensions, emulsions, microemulsions,micellar solutions, gels, and ointments. The pharmaceutically acceptablecarriers may also contain ingredients that include, but are not limitedto, saline and aqueous electrolyte solutions; ionic and nonionic osmoticagents such as sodium chloride, potassium chloride, glycerol, anddextrose; pH adjusters and buffers such as salts of hydroxide,phosphate, citrate, acetate, borate; and trolamine; antioxidants such assalts, acids and/or bases of bisulfite, sulfite, metabisulfite,thiosulfite, ascorbic acid, acetyl cysteine, cysteine, glutathione,butylated hydroxyanisole, butylated hydroxytoluene, tocopherols, andascorbyl palmitate; surfactants such as lecithin, phospholipids,including but not limited to phosphatidylcholine,phosphatidylethanolamine and phosphatidyl inositiol; poloxamers andpoloxamines, polysorbates such as polysorbate 80, polysorbate 60, andpolysorbate 20, polyethers such as polyethylene glycols andpolypropylene glycols; polyvinyls such as polyvinyl alcohol andpovidone; cellulose derivatives such as methylcellulose, hydroxypropylcellulose, hydroxyethyl cellulose, carboxymethyl cellulose andhydroxypropyl methylcellulose and their salts; petroleum derivativessuch as mineral oil and white petrolatum; fats such as lanolin, peanutoil, palm oil, soybean oil; mono-, di-, and triglycerides; polymers ofacrylic acid such as carboxypolymethylene gel, and hydrophobicallymodified cross-linked acrylate copolymer; polysaccharides such asdextrans and glycosaminoglycans such as sodium hyaluronate. Suchpharmaceutically acceptable carriers may be preserved against bacterialcontamination using well-known preservatives, these include, but are notlimited to, benzalkonium chloride, ethylenediaminetetraacetic acid andits salts, benzethonium chloride, chlorhexidine, chlorobutanol,methylparaben, thimerosal, and phenylethyl alcohol, or may be formulatedas a non-preserved formulation for either single or multiple use.

For example, a tablet formulation or a capsule formulation of the activecompound may contain other excipients that have no bioactivity and noreaction with the active compound. Excipients of a tablet or a capsulemay include fillers, binders, lubricants and glidants, disintegrators,wetting agents, and release rate modifiers. Binders promote the adhesionof particles of the formulation and are important for a tabletformulation. Examples of excipients of a tablet or a capsule include,but not limited to, carboxymethylcellulose, cellulose, ethylcellulose,hydroxypropylmethylcellulose, methylcellulose, karaya gum, starch,tragacanth gum, gelatin, magnesium stearate, titanium dioxide,poly(acrylic acid), and polyvinylpyrrolidone. For example, a tabletformulation may contain inactive ingredients such as colloidal silicondioxide, crospovidone, hypromellose, magnesium stearate,microcrystalline cellulose, polyethylene glycol, sodium starchglycolate, and/or titanium dioxide. A capsule formulation may containinactive ingredients such as gelatin, magnesium stearate, and/ortitanium dioxide.

For example, a patch formulation of the active compound may comprisesome inactive ingredients such as 1,3-butylene glycol, dihydroxyaluminumaminoacetate, disodium edetate, D-sorbitol, gelatin, kaolin,methylparaben, polysorbate 80, povidone, propylene glycol,propylparaben, sodium carboxymethylcellulose, sodium polyacrylate,tartaric acid, titanium dioxide, and purified water. A patch formulationmay also contain skin permeability enhancer such as lactate esters(e.g., lauryl lactate) or diethylene glycol monoethyl ether.

Topical formulations including the active compound can be in a form ofgel, cream, lotion, liquid, emulsion, ointment, spray, solution, andsuspension. The inactive ingredients in the topical formulations forexample include, but not limited to, lauryl lactate(emollient/permeation enhancer), diethylene glycol monoethyl ether(emollient/permeation enhancer), DMSO (solubility enhancer), siliconeelastomer (rheology/texture modifier), caprylic/capric triglyceride,(emollient), octisalate, (emollient/UV filter), silicone fluid(emollient/diluent), squalene (emollient), sunflower oil (emollient),and silicone dioxide (thickening agent).

In one embodiment, DMSO at 0.1-20%, or 0.5-10% (w/w) is included in thetopical gel formulation, which provides suitable solubility of theactive compound.

In another embodiment, diethylene glycol monoethyl ether is included inthe topical gel formulation.

Method of Use

Inflammation is a process and a state of tissue pathology resulting fromactivation and continuation of activity of the innate and acquiredcomponents of the immune system. The arachidonic acid cascade andcytokine production and action in cell to cell interactions are criticalcomponents of immune activation and response, which lead toinflammation. Arachidonic acid is a component of membrane phospholipids.After it is freed from phospholipids, arachidonic acid acts as aprecursor to many of the known eicosanoids including prostaglandins andleukotrienes, which are known pro-inflammatory entities.

The active compound is anti-inflammatory when applied topically in themouse ear swelling model, in which the inflammation is induced byarachidonic acid. The active compound is effective in inhibitingpro-inflammatory mediators.

The present invention is directed to a method of treating inflammationand/or pain. 4-methylsulfonyl-2-butenenitrile, can be used as is, or itcan be administered in the form of a pharmaceutical composition thatadditionally contains a pharmaceutically acceptable carrier. The methodcomprises the steps of first identifying a subject suffering frominflammation and/or pain, and administering to the subject the activecompound, in an amount effective to treat inflammation and/or pain. “Aneffective amount,” as used herein, is the amount effective to treat adisease by ameliorating the pathological condition or reducing thesymptoms of the disease.

In one embodiment, the method reduces or alleviates the symptomsassociated with inflammation. The present invention provides a method totreat localized manifestations of inflammation characterized by acute orchronic swelling, pain, redness, increased temperature, or loss offunction in some cases.

In another embodiment, the present invention provides a method toalleviate the symptoms of pain regardless of the cause of the pain. Thegeneral term “pain” treatable by the present method includesnociceptive, neuropathic, and mix-type. The present invention reducespain of varying severity, i.e. mild, moderate and severe pain; acute andchronic pain. The present invention is effective in treating joint pain,muscle pain, tendon pain, burn pain, and pain caused by inflammationsuch as rheumatoid arthritis.

In one embodiment, the present invention is useful in treatinginflammation and/or pain associated in a musculoskeletal system or onthe skin. The highly innervated, musculoskeletal and skin systems have ahigh capacity for demonstration of pain. In addition, themusculoskeletal system has a high capacity for tissue swelling, and theskin has a high capacity for redness, swelling, and heat. Inmusculoskeletal and skin systems, the degree of tissue damage isfrequently magnified out of proportion to the resulting inflammatoryresponse. In the skin for example, merely firm stroking will causerelease of the cytokines, IL-1 and TNF.

The present invention provides a method for treating inflammation and/orpain associated with inflammatory skeletal or muscular diseases orconditions. The method comprises the steps of identifying a subject inneed thereof, and administering to the subject the active compound, inan amount effective to treat inflammation and/or pain. “The activecompound,” as used in this application, is intended to include4-methylsulfonyl-2-butenenitrile and its pharmaceutically acceptablesalts, isomer, or isomers thereof. The skeletal or muscular diseases orconditions include musculoskeletal sprains, musculoskeletal strains,tendonopathy, peripheral radiculopathy, osteoarthritis, jointdegenerative disease, polymyalgia rheumatica, juvenile arthritis, gout,ankylosing spondylitis, psoriatic arthritis, systemic lupuserythematosus, costochondritis, tendonitis, bursitis, such as the commonlateral epicondylitis (tennis elbow), medial epicondylitis (pitcherselbow) and trochanteric bursitis, temporomandibular joint syndrome, andfibromyalgia.

The present invention provides a method for treating inflammation and/orpain associated with inflammatory skin diseases such as dermatitis,psoriasis, and acne. The method comprises the steps of identifying asubject in need thereof, and administering to the subject the activecompound, in an amount effective to treat inflammation and/or pain.

Skin is highly reactive to environmental stimuli and the epidermalcomponent of keratinocytes is a very rich source of both arachidonicacid and pro-inflammatory cytokines of IL-1 and TNF. The skin dendriticcells, Langerhans cells, recognize and process antigens for furtherimmune response of various lymphocytes and all of these cells areprimarily regulated by cytokines through their specific cell surfacereceptors.

4-Methylsulfonyl-2-butenenitrile, which is effective in inhibitingarachidonic acid induced inflammation and inhibiting pro-inflammatorymediators, is effective to treat inflammation and/or pain associatedwith psoriasis, acne, rosacea, and dermatitis, particularly contactdermatitis, and atopic dermatitis. The present invention provides amethod for treating inflammation and/or pain associated withinflammatory skin diseases such as psoriasis, acne, rosacea, anddermatitis, particularly contact dermatitis, and atopic dermatitis. Themethod comprises the steps of identifying a subject in need thereof, andadministering to the subject the active compound, in an amount effectiveto treat inflammation and/or pain.

4-Methylsulfonyl-2-butenenitrile, which are effective in inhibitingarachidonic acid induced inflammation and in inhibiting the release ofpro-inflammatory cytokine, are effective to treat inflammatory skindiseases such as dermatitis (atopic dermatitis), psoriasis, acne, androsacea. The present invention provides a method for treatinginflammatory skin diseases such as dermatitis, psoriasis, and acne (acnevulgaris). The method comprises the steps of identifying a subject inneed thereof, and administering to the subject4-methylsulfonyl-2-butenenitrile, or a pharmaceutically acceptable saltthereof, in an amount effective to reduce or eliminate the symptoms ofthe disease.

Dermatitis (also called eczema) is generic inflammation of the skin.Specific types of dermatitis include atopic, contact, nummular, andphoto-induced.

Contact dermatitis is a localized rash or irritation of the skin causedby contact with a foreign substance. Only the superficial regions of theskin are affected in contact dermatitis. Inflammation of the affectedtissue is present in the epidermis (the outermost layer of skin) and theouter dermis (the layer beneath the epidermis). Contact dermatitisresults in large, burning, and itchy rashes. Contact dermatitis is aninflammatory condition of the skin either of irritant exposure to theskin without specific adaptive immunologic pathogenesis or of allergicsensitization and subsequent exposure of the skin to the sensitizingallergen with specific adaptive immunologic pathogenesis. Both involveinnate and acquired immune system response including arachidonic acidand cytokine components that initiate and propagate the disease throughcell to cell messaging by eicosanoid and/or cytokine moieties producedby epidermal cells, macrophages, dendritic cells, neutrophils,eosinophils, and various T and B lymphocytes. Contact dermatitis may beeither acute or chronic. The acute forms are pruritic with erythema,edema, and micro or macrovesiculation in the areas of skin contact bythe initiating factor. The chronic forms are pruritic with mildererythema, scaling, lichenification, and possibly fissuring particularlyon the hands.

Allergic contact dermatitis is a T cell-mediated delayed typehypersensitivity reaction that occurs upon hapten challenge insensitized individuals. The inflammatory response in classical allergiccontact dermatitis requires both a sensitization phase and anelicitation phase responsible for the recruitment and activation ofspecific T cells at the site of hapten skin challenge.

Atopic dermatitis is a genetically determined disease that is part ofthe broader disease complex of atopy that includes asthma, hay fever,and atopic dermatitis. Many individuals with atopic dermatitis havevarious mutations of the filaggrin gene that codes for an importantepidermal structural protein that when defective, results in abnormalbarrier function of the epidermis. The altered barrier allows exposureto multiple environmental allergens that are first recognized by innateimmune responses involving arachidonic acid and eicosanoids andrecruitment of eosinophils, mast cells, and other inflammatory cellsthat initiate an acute responses of itch, erythema, and subsequentscratching and additionally activate the adaptive immune responses thatinvolve inflammation by lymphocytes predominantly of a TH 2 derivationand activity. Atopic dermatitis is responsive to a number of cytokineinhibitors such as cyclosporine, and tacrolimus.

Current theory of the pathogenesis of psoriasis is that in individualswho are genetically susceptible a triggering event in the epidermis suchas injury or super antigen contact initiates an response of the innateimmune system with arachidonic acid and eicosanoid generation,recruitment and activity of neutrophils. Subsequent transformation ofthe response to that of a TH 1 adaptive immunity with cytokineactivation and activity of specific T lymphocytes effect thepathological changes in the epidermis and dermis, which result in thetypical psoriasis lesions of plaques that are erythematous, thickened,and scaly. Psoriasis is responsive to various immunomodulators includingcyclosporine, methotrexate, and a host of specific biologicals thatinterfere with cytokine signaling.

Acne vulgaris, a progressively inflammatory disorder of thepilosebaceous follicular unit especially of the face and upper chest andback is a very common disease of both males and females after initiationof puberty, and in females even prior to adrenal gland maturity.Increased production of androgenic hormones by adrenal, ovarian, andtesticular glands and by the pilosebaceous unit itself produce anincrease in sebum and changes in its lipid composition, which combinewith follicular epithelial cells to produce some degree of obstructionof the infra-infundibular portion of the pilosebaceous follicleresulting in the initial lesion of acne, the microcomedo. Thisconsequent dilation of the pore and the changed composition of sebum atpuberty facilitate colonization of the follicle by Propionibacteriumacnes bacilli that produce enzymes to degrade the triglycerides in sebumto free fatty acids that leak through the follicle into the dermis andincite arachidonic acid pathways of eicosanoid production and subsequentinitiation of inflammation. The bacilli also initiate chemokineproduction that attracts further inflammatory cells to the area andconsequent cytokine production and action to continue and amplifyinflammation. Thus initiation and propagation of progressiveinflammation in the microcomedo produces the evolution to the severalhallmark lesions of inflammatory acne, papule, pustule, nodule, andcyst. The present invention is useful to treat common acne, comedonicacne, papulopustular acne, papulocomedonic acne, nodulocystic acne, acneconglobata, cheloid acne of the nape of the neck, recurrent miliaryacne, necrotic acne, neonatal acne, occupational acne, acne rosacea,senile acne, solar acne or acne medicamentosa.

Rosacea is a chronic condition characterized by facial erythema andsometimes pimples. Rosacea typically begins as redness on the centralface across the cheeks, nose, or forehead, but can also less commonlyaffect the neck, chest, ears, and scalp. In some cases, additionalsymptoms, such as semi-permanent redness, telangiectasia (dilation ofsuperficial blood vessels on the face), red domed papules (small bumps)and pustules, red gritty eyes, burning and stinging sensations, and insome advanced cases, a red lobulated nose (rhinophyma), may develop.There are 3 subtypes of rosacea that affect the skin:erythematotelangiectatic rosacea, papulopustular rosacea, and phymatousrosacea.

4-Methylsulfonyl-2-butenenitrile are effective in treating contactdermatitis and alleviating one or more symptoms selected from the groupconsisting of consisting of erythema, edema, lichenification, scaling,fissuring, and micro or macrovesiculation.

4-Methylsulfonyl-2-butenenitrile are effective in treating atopicdermatitis and alleviating one or more symptoms selected from the groupconsisting of erythema, induration, lichenification, scaling, and oozingand crusting.

4-Methylsulfonyl-2-butenenitrile are effective in treating psoriasis andalleviating erythema, scaling, and/or thickness of the psoriasislesions.

4-Methylsulfonyl-2-butenenitrile are effective in treating acne andalleviating acne lesions selected from the groups consisting of closedcomedones, papules, pustules, nodules, and cysts.

4-Methylsulfonyl-2-butenenitrile are effective in treating rosacea andalleviating one or more symptoms selected from the group consisting oferythema, telangiectasia, red domed papules and pustules, red grittyeyes, and burning and stinging sensations.

The pharmaceutical composition of the present invention can be appliedby local administration and systemic administration. Localadministration includes topical administration. Systemic administrationincludes oral, parenteral (such as intravenous, intramuscular,subcutaneous or rectal), and other systemic routes of administration. Insystemic administration, the active compound first reaches plasma andthen distributes into target tissues. Topical administration and oraladministration are preferred routes of administration for the presentinvention.

Dosing of the composition can vary based on the extent of the injury andeach patient's individual response. For systemic administration, plasmaconcentrations of the active compound delivered can vary; but aregenerally 1×10⁻¹⁰-1×10⁻⁴ moles/liter, and preferably 1×10⁻⁸-1×10⁻⁵moles/liter.

In one embodiment, the composition is applied topically onto theaffected area and rubbed into it. The composition is topically appliedat least 1 or 2 times a day, or 3 to 4 times per day, depending on themedical issue and the disease pathology being chronic or acute. Ingeneral, the topical composition comprises about 0.01-20%, or 0.05-20%,or 0.1-20%, or 0.2-15%, 0.5-10, or 1-5% (w/w) of the active compound.For example, the topical composition comprises about 1 or 5% (w/w) ofthe active compound. Depending on the size of the affected area, 0.2-85mL, typically 0.2-10 mL, of the topical composition is applied to theindividual per dose. The active compound passes through skin and isdelivered to the site of discomfort.

In one embodiment, the pharmaceutical composition is administratedorally to the subject. The dosage for oral administration is generallyat least 0.1 mg/kg/day and less than 100 mg/kg/day. For example, thedosage for oral administration is 0.1-100, or 0.5-50, or 1-20, or 1-10,or 1-50 mg/kg/day for a human subject. For example, the dosage for oraladministration is 20-1000 mg/day, and preferably 20-500, 20-100, 25-200,50-500, 50-200, 100-600, 100-400, or 200-800 mg/day for a human subject.

In one embodiment, the pharmaceutical composition is administratedintravenously to the subject. The dosage for intravenous bolus injectionor intravenous infusion is generally 0.03 to 20 and preferably 0.03 to10 mg/kg/day.

In one embodiment, the pharmaceutical composition is administratedsubcutaneously to the subject. The dosage for subcutaneousadministration is generally 0.3-20, and preferably 0.3-3 mg/kg/day.

Those of skill in the art will recognize that a wide variety of deliverymechanisms are also suitable for the present invention.

The present invention is useful in treating a mammal subject, such ashumans, horses, and dogs. The present invention is particularly usefulin treating humans.

The following examples further illustrate the present invention. Theseexamples are intended merely to be illustrative of the present inventionand are not to be construed as being limiting.

Examples Example 1. Preparation of 4-methylsulfonyl-2-butenenitrile

(A) A solution of allyl cyanide (0.59 mole) in t-amyl alcohol (120 mL)and petroleum ether (370 mL) was treated sequentially with a solution ofbromine (0.59 mole) in t-amyl alcohol (60 mL), followed by an ethanolicsolution of sodium ethoxide (345 mL, 0.60 mole). When the reactionmixture had cooled to RT, solids were removed by vacuum filtration andthe filtrate was concentrated under reduced pressure. The residualliquid was charged on a silica gel column (307.10 g) and eluted withhexanes-ethyl acetate (19:1; 9:1). Combination of appropriate fractions,followed by concentration under reduced pressure gave4-bromo-2-butenenitrile as a pale yellow liquid (47.14 g).

(B) The product from part (A) (150 mmol) was added dropwise over ˜14 minto an equimolar solution of sodium methanesulfinate in ethanol (120 mL)containing water (4 mL) stirred at ˜40° C. After continued heating andstirring for 6 h, the reaction mixture was concentrated under reducedpressure. The dark residual syrup was diluted with acetone, then vacuumfiltered to remove sodium bromide. The filtrate was concentrated to adark brown syrup, which was charged on a silica gel column (˜255 g) thatwas eluted with chloroform-ethanol (19:1). Combination of appropriatefractions, followed by concentration under reduced pressure gave a lightamber-colored liquid (16.08 g). The liquid (10.48 g) was charged on asilica gel column (˜310 g) that was eluted withchloroform-ethanol-hexane (66:4:30). Combination of appropriatefractions, followed by concentration under reduced pressure gave a lightamber-colored liquid, which was taken up in acetone and passed through a0.45 μm syringe filter to remove a fine particulate. Concentration ofthe clear filtrate gave the expected product (9.71 g). The ¹H NMRspectrum for 4-methylsulfonyl-2-butenenitrile was acquired in DMSO-d₆solution at 400 MHz by Spectral Data Services, Inc. Table 1 shows thechemical shift data (ppm) of NMR results.

TABLE 1 ¹H Chemical shifts (ppm) for 4-methylsulfonyl-2-butenenitrile.Num- Chem- ber ical of Shift Pro- (ppm) RIV ^(a) tons MultiplicityAssignment 1.24-1.28 0.11 impurity 1.32-1.36 0.10 impurity 2.09 3.03acetone 2.32-2.36 0.07 impurity 2.50 DMSO 3.0-3.2 73.85 3 d CH₃SO₂3.2-3.3 0.23 impurity 3.5-4.2 1.22 impurities (4 signals) 4.1-4.3 47.982 dd CH₃SO₂CH ₂ 4.3-4.4 0.39 impurity 5.2-5.3 0.18 impurity 5.72-5.960.49 impurities (multiple signals) 6.08 ~9.5 1 dm (J ~16 Hz)SO₂CH₂CH═CHCN (E) 6.12 ~13 d (J ~7 Hz) SO₂CH₂CH═CHCN (Z) 6.2-6.6 0.47impurities (multiple signals) 6.64-6.92 23.31 1 dm SO₂CH₂CH═CHCN (E,Z)6.92-7.12 0.49 impurities (multiple signals)

H NMR: δ 3.0-3.2 (d, 3H), 4.1-4.3 (dd, 2H), [6.08 (dm, J˜16 Hz)+6.12 (d,J˜7 Hz), 1H], 6.64-6.92 (dm, 1H).

The structure of 4-methylsulfonyl-2-butenenitrile comprises 7 protons.All protons are accounted for and occur at reasonable chemical shifts.Chemical shift assignments for the methyl and methylene protons areunequivocal. The two sets of multiplets between 6 ppm and 6.2 ppm can bedistinguished by their coupling constants (J). J˜16 Hz is consistentwith the coupling constant expected for alkene protons in anE-configuration, whereas J˜11 Hz is consistent with the couplingconstant expected for alkene protons in a Z-configuration. Based on acomparison of RIV values for these signals, the E/Z ratio is ˜58/42.Coupling constants for the remaining alkene protons cannot be measuredexactly due to the complexity of the signals between 6.6 ppm and 7 ppm.The spectrum contains some impurity/solvent peaks (RIV_(Total) 3.75).

Example 2. Gel Formulation 1

Table 2 exemplifies one gel formulation containing4-methylsulfonyl-2-butenenitrile.

TABLE 2 1% Gel 5% Gel Active compound 1.0% 5.0% Diethylene glycolmonoethyl ether 5.0% 5.0% Acrylates/C10-30 alkyl acrylate 0.75% 0.75%crosspolymer (CARBOPOL ® Ultrez 20 polymer) Trolamine(tris(2-hydroxyethyl)amine) 1.13% 1.13% Parabens 0.07% 0.07% Propyleneglycol 0.43% 0.43% Purified Water 91.62% 87.62% Total 100.0% 100.0%

Example 3. Anti-Inflammatory Activity of Active Compound by TopicalApplication in Mice

4-Methylsulfonyl-2-butenenitrile, prepared from Example 1, was used inthis experiment.

The test compound, indomethacin (positive control), and vehicle wereevaluated for anti-inflammatory activity in a topical arachidonicacid-induced ear swelling model in mice.

Male ICR mice weighing 22±2 g were used and randomly divided; the testcompound and vehicle control had 10 mice, and indomethacin had 5 mice.Arachidonic Acid (0.5 mg in 20 μl of acetone:ethanol/1:1) was appliedtopically to the anterior and posterior surfaces of the right ear ofeach mouse. Test substances and vehicle, as listed in Table 2 weresimilarly applied 30 min before and 15 min after arachidonic acidapplication. The thickness of the right ear and the left ear wasmeasured and the difference calculated as an indication of theinflammation in the right ear. Ear swelling was measured by a Dyer modelmicrometer gauge at 60 and 90 minutes after arachidonic acid applicationas an index of inflammation. Percent inhibition was calculated accordingto the formula: Ic−It/Ic×100, where Ic and It refers to increase of earthickness (mm) in control and treated mice, respectively. ANOVA andDunnett's test were employed to ascertain significant difference betweenvehicle control and treated groups. Significance is set at P<0.05 level.The results measured at 90 minutes after arachidonic acid applicationare summarized in Table 3.

TABLE 3 Dosage Conc mg/20 % Test Substance mM μL n Inhibition P ValueVehicle - acetone:ethanol NA NA 10 NA (1:1) Indomethacin 14 0.1 5 58<0.001 (Positive control) 4-methylsulfonyl-2- 375 1.1 10 22 0.002butenenitrile

The tested compound resulted in 22% inhibition in the ear swellinginduced by arachidonic acid, relative to that in the vehicle-treatedgroup. The differences between treated mice and vehicle-treated micewere determined to be statistically significant (p-value by t-test was0.002).

Example 4. Anti-Inflammatory Activity of Active Compound in Mice by OralApplication (Prophetic Example)

The active compound 4-methylsulfonyl-2-butenenitrile is dissolved in avehicle (1% Tween 80 in water suspension or other lipid-basedpharmaceutically acceptable carrier) to 5-15 mg/mL. The test compound,dexamethasone (positive control), and vehicle are orally administered tomice and evaluated for anti-inflammatory activity in the topicalarachidonic acid induced ear swelling model in mice.

Male ICR derived mice are used in this experiment. 10 mice are used foreach group (active compound, positive control, and vehicle). All animalsare maintained in a controlled temperature (22-24° C.) and humidity(60%-70%) environment with 12-hour light/dark cycles for at least oneweek prior to use.

Arachidonic acid (0.5 mg in 20 μL acetone) is applied topically onto theanterior and posterior surfaces of the right ear of test animals toinduce inflammation. Test compound in vehicle (10 mL/kg) and vehicle (10mL/kg, 50-150 mg/kg) are orally administered by gavage 1 hour beforearachidonic acid, whereas dexamethasone is orally administered by gavage3 hour before arachidonic acid challenge. At 60 minutes and 90 minutesafter arachidonic acid induction of ear edema, the thickness of theright ear and the left ear is measured and the difference calculated asan indication of the inflammation in the right ear. Significant activityis defined as a statistically significant inhibition (p-value determinedby t-test was <0.05) in arachidonic acid induced ear swelling relativeto the vehicle-treated group.

Example 5. Analgesic Activity of Active Compound by Oral Administrationin Mice (Formalin Model, Prophetic Example)

Formalin test is a model of continuous pain resulting fromformalin-induced tissue injury. Nociceptive and inflammatory pain isinduced by injection of a dilute formalin solution into the paw,resulting in nocifensive behavior including paw flinching. The formalinmodel encompasses inflammatory, neurogenic, and central mechanism ofpain. The early phase of pain (from 0 to about 10 minutes) is due tonociceptive mechanism and the late phase of pain (from 10-40 minutes) isdue to a combination of inflammatory pain and nociceptive mechanism.Pain behavior is assessed using manual paw licking measurements. Theendpoints of the study are the number of paw licking events. (Hunskaaret al., Pain, 30:103-114, 1987; Li et al., Molecular Pain, 6:11, 2010)

10 Mice per group are used in the study. Immediately prior to testing(at time 0), mice are restrained in a cloth and injected with 20 μL of a5% formalin solution, subcutaneously into the dorsal surface of the lefthind paw. Vehicle control (DMSO) and test compound4-methylsulfonyl-2-butenenitrile (in DMSO) are administered by oralgavage with a volume of 5 mL/kg to mice. The amounts of test compoundare 100 or 500 mg/kg per dose.

Positive control morphine in saline is administered by subcutaneousinjection at 8 mg/kg to mice, immediately before formalin injection andtesting at time zero.

Following formalin injection, animals are placed in individual cages,and manually observed for 60 minutes. The licking events are recorded infive minute intervals continuously for a total of 60 minutes.

The number of licking events at different time points post formalininjection of vehicle control, morphine-treated, and testcompound-treated mice are plotted in 5 minute intervals.

The numbers of licking events per minute are calculated between 0-10minutes and 10-40 minutes for vehicle, positive control, and testcompound. A statistically significant reduction of licking event perminute is an indication that the test compound is effective in treatingacute nociceptive pain (early phase) or inflammatory nociceptive pain(late phase).

Example 6. Analgesic Activity of Active Compound by TopicalAdministration in Mice (Formalin Model, Prophetic Example)

The animals and the treatment protocol are similar to those described inExample 5, except the following.

The test compound 4-methylsulfonyl-2-butenenitrile (375 mM in vehicle,n=10) and vehicle control (acetone:ethanol 1:1, n=10) are administeredtopically by submerging the mouse left hind paw in the respectivesolution for about 30 seconds. The paw is then withdrawn and wiped withtissue to avoid excess dermal drying.

Positive control morphine is administered by subcutaneous injection at 8mg/kg in saline (n=10).

Morphine is subcutaneously administered once 15 minutes before formalininjection. The test compounds and vehicle control are topicallyadministered twice (BID), at 60 minutes before formalin injection.

Following formalin injection, animals are placed in individual cages,and manually observed for 60 minutes. The licking events are recorded infive minute intervals continuously for a total of 40 minutes.

The number of licking events at different time points post formalininjection of vehicle control, morphine-treated, and testcompound-treated mice are determined.

The numbers of licking events per minute are calculated between 0-10minutes and 10-40 minutes for vehicle, positive control, and testcompound. A two-sample t-test is done to compare the vehicle group withthe test compound group. Significance is set at P<0.05 level.

Example 7. Analgesic Activity of Active Compound in Chronic ConstrictionInjury Model (Prophetic Example)

Peripheral nerve lesions may generate a syndrome comprising, in additionto spontaneous pain, exaggerated responses to light touch (tactileallodynia). Chronic constriction injury model is a neuropathic painmodel.

Male Sprague Dawley rats are used. Under pentobarbital (50 mg/kg, 5ml/kg, i.p.) anesthesia, the sciatic nerve is exposed at mid-thighlevel. Four ligatures (4-0 chromic gut), about 1 mm apart, are looselytied around the nerve. The animals are then housed individually in cageswith soft bedding for 7 days before testing. Constriction of the sciaticnerve produces nerve injury and unilateral neuropathic pain.

On the day of experiments, the animals have no access to food overnightbefore testing. The rats are placed under inverted plexiglass cages on awire mesh rack and allowed to acclimate for 20 to 30 minutes. Mechanicallodynia is evaluated by the Chaplan up/down method using von Freyfilaments to the plantar surface of the left hind paw. See Chaplan, etal. J. Neuroscience Methods, 53: 55-63, 1994.

Rats are pre-selected for experimentation only if the pain threshold7-14 days after nerve ligation (pre-treatment) is reduced by 10 grams offorce relative to the response of the individual paw before nerveligation (pre-ligation), namely, with clear presence of allodynia.

The active compound 4-methylsulfonyl-2-butenenitrile is prepared in thegel formulation according to Example 2.

Active compound in gel formulation (1 or 5%, topical administration),active compound in DMSO (500 mg/kg, oral administration), morphine(positive control, subcutaneous, 8 mg/kg in saline), topical vehicle(gel formulation without the active compound), and oral vehicle (DMSO)are evaluated.

Test substance or vehicle is either administered orally or topically tothe plantar surface of the left hind paw. The mechanical allodynia testis performed 30 min before (pre-treatment) and 1 and 3 hours after asingle dose of test substance or vehicle (post treatment). Paw withdrawthresholds of control and tested compound are measured.

Example 8. Treatment of Knee Pain by Topical Administration (PropheticExample)

Objectives:

To investigate the efficacy of the active compound in a topical gelformulation in human patients with mild to severe knee pain associatedwith osteoarthritis following temporary cessation of standard NSAIDtherapy. The focus of this study is on the symptoms caused by painfularthritis. The clinical trial is utilizing osteoarthritis of the knee asa well-established paradigm for other musculoskeletal disorders.

Topical Formulation:

The gel formulations containing the active compound4-methylsulfonyl-2-butenenitrile at 1% and 5% (Example 2) are used inthis example. Placebo contains the same gel without the active compound.

Methodology:

A randomized, double-blind, placebo controlled, parallel treatmentmulticenter clinical activity study.

Patients with painful osteoarthritis of the knee, controlled by a stabledose of standard NSAID therapy for at least 2 months, discontinue use ofthe NSAIDs for a 7-day washout period. Patients are then randomized in a1:1:1 ratio (1% active gel, 5% active gel, placebo). A total of up to150 patients are enrolled.

The active gel or placebo is applied to the affected knee 3 times a dayfor 12 weeks for a total of 252 treatments given every 4-6 hours whileawake.

Patients are treated for 12 weeks and followed up for a further 4 weeks.NSAIDs may be restarted after the Week 12 visit.

Criteria for Evaluation:

Safety:

-   -   Adverse Events (AEs) throughout the study.    -   Physical examination at enrollment (˜7 days, start of NSAID        washout period), Baseline (Day 1, start of treatment), Week 12        and Week 16.    -   Vital signs at enrollment (−7 days, start of NSAID washout        period), Baseline (Day 1, start of treatment) and Weeks 2, 4, 6,        12 and 16.    -   Clinical laboratory measurements at Baseline (Day 1), Week 4, 8,        12 and 16.

Clinical Activity:

The primary clinical activity parameters are the measurement of pain inthe target joint, as quantified by the Visual Analog Scale (VAS) and theWestern Ontario and McMaster University (WOMAC) Index pain subscale. Theeffect of treatment on swelling, tenderness and inflammation of the kneeis recorded, also the time to reduction or eradication of pain aftertreatment is recorded.

Study Endpoints:

The primary clinical activity endpoint is:

-   -   Change from Baseline (Day 1) to Week 12 in WOMAC functional        disability index pain subscale (Scale 0-20)

The secondary clinical activity endpoints are:

-   -   Change from Baseline (Day 1) to Week 12 in WOMAC functional        disability index subscales:    -   Stiffness (Scale 0-8).    -   Physical function (Scale 0-68).    -   Change from Baseline (Day 1) to Week 12 in VAS pain score        (0-100).    -   Change from Baseline (Day 1) to Week 2 in VAS pain score        (0-100).    -   Change in investigator evaluation of swelling, tenderness and        inflammation between Baseline (Day 1) and Weeks 4 and 12 after        the first application on Day 1.    -   Time to reduction or eradication of pain subsequent to each        topical application of active gel or placebo gel.    -   Use of rescue medication (APAP).

Example 9. Treatment of Knee Pain by Oral Administration (PropheticExample)

The design and protocols of this experiment are similar to thosedescribed in Example 9, except the active compounds and placebo areapplied by an oral route.

Oral Formulation:

Tablet formulations containing 10, 100, or 1000 mg of the activecompound 4-methylsulfonyl-2-butenenitrile are used in this example.Placebo has the same tablet formulation without the active compound.

Methodology:

Patients are then randomized in a 1:1:1:1 ratio (10 mg:100 mg:1000mg:placebo). A total of up to 200 patients are enrolled.

The active tablet or placebo is administered orally to each patient twotimes a day for 12 weeks for a total of 168 treatments given every 12hours while awake. Patients are treated for 12 weeks and followed up fora further 4 weeks.

Criteria for evaluation are the same as those described in Example 9.

Example 10. Treatment of Arthritis (Prophetic Example)

Zymosan injected directly into the knee joint of mice elicits aninflammatory response and is used as a model of arthritis (Verschure etal, Ann. Rheum Dis. 53:455-460, 1994). Endpoints measured in this modelinclude knee joint swelling score, cytokine levels in the synovialtissue and microscopic pathology of the knee.

Active compound 4-methylsulfonyl-2-butenenitrile (in vehicle) andvehicle control (1% Tween 80 in water suspension or other lipid-basedpharmaceutically acceptable carrier) are administered by oral gavage tomice with a volume of 5 mL/kg. The oral dosage of active compound is 500mg/kg mouse.

There are 5 mice per group, with a total of 10 knees injected. On Day 1,C57BL6mice are dosed (500 mg/kg) with active compound or vehicle twiceon Hours 0 and 12. On Day 2, mice are dosed with active compound orvehicle on Hour 24, then injected intra-articularly with 180 μg ofzymosan (6 μL) into both knee joints on Hour 25, and then dosed a secondtime on Hour 36 with each active compound or vehicle. On Day 3, mice areagain dosed with active compound or vehicle on Hour 48. Two hourpost-dosing on Hour 50, knees are scored for edema, synovial tissue iscollected for measurement of cytokine levels, and knee joints areprocessed for histopathology for analysis of inflammatory immune cellinflux into the joint. Macroscopic joint swelling is assessed on allknees after the skin is removed using a scoring system ranging from 0 to3, with 0 being no swelling and 3 being severe swelling. Synovial tissueis taken from 5 knees for measurement of mouse interleukin-10,interleukin-6, and interleukin-1 receptor antagonist levels. Theremaining 5 knees are processed for microscopic pathology for assessmentof cellular influx into the site of inflammation.

Results for each group are presented as mean±standard error of mean andstatistical evaluation is performed.

Treatment with active compound is expected to result in decreasedinflammation as measured by a decrease in joint swelling, decrease incytokine levels and decrease influx of inflammatory cells to the site ofinflammation.

Example 11. Treatment of Gout (Prophetic Example)

Monosodium urate monohydrate (MSU) crystals injected in combination witha free fatty acid (FFA) directly into the knee joint of mice elicits aninflammatory response and is used as a model of gout (Joosten et al,Arthritis & Rheumatism, 62:3237-3248, 2010). Endpoints measured in thismodel include knee joint swelling score, cytokine levels in the synovialtissue and microscopic pathology of the knee.

Active compound 4-methylsulfonyl-2-butenenitrile (in vehicle) andvehicle control (1% Tween 80 in water suspension or other lipid-basedpharmaceutically acceptable carrier) are administered by oral gavage tomice with a volume of 5 mL/kg. The oral dosage of active compound is 500mg/kg mouse.

There are 5 mice per group, with a total of 10 knees injected. On Day 1,C57BL6 mice are dosed (500 mg/kg/dose) with active compounds or vehicletwice on Hours 0 and 12. On Day 2, mice are dosed with active compoundsor vehicle on Hour 24, then injected intra-articularly with MSU crystals(300 μg) and C18:0 FFA (200 μM, 10 μL) on Hour 25. Three hours later(Hour 28), knees are scored for edema, synovial tissue is collected formeasurement of cytokine levels, and knee joints are processed forhistopathology for analysis of inflammatory immune cell influx into thejoint. Macroscopic joint swelling is assessed on all knees after theskin is removed using a scoring system ranging from 0 to 3, with 0 beingno swelling and 3 being severe swelling. Synovial tissue is taken from 5knees for measurement of mouse interleukin-1β, interleukin-6, andinterleukin-1 receptor antagonist levels. The remaining 5 knees areprocessed for microscopic pathology for assessment of cellular influxinto the site of inflammation.

Results for each group are presented as mean±standard error of mean andstatistical evaluation is performed.

Treatment with active compound is expected to result in decreasedinflammation as measured by a decrease in joint swelling, decrease incytokine levels and decrease influx of inflammatory cells to the site ofinflammation.

Example 12. Treatment of Contact Dermatitis (Prophetic Example)

Mice dermally sensitized and challenged by dinitrofluorobenzene (DNFB)are used as a model of contact dermatitis (Saint-Mezard, J InvestDermatol, 120:641-647, 2003).

Sensitization and Challenging:

There are 5 mice per group. Each mouse is sensitized with 0.5% DNFB(vehicle=4:1 (vol/vol) acetone:olive oil) topically on the shavedabdomen, 6 days before challenge. The right ears of the mice are thenchallenged with a topical application of 0.2% DNFB in vehicle. The leftears of the mice receive the vehicle as control.

Oral Administration:

Active compound 4-methylsulfonyl-2-butenenitrile (in vehicle) andvehicle control (1% Tween 80 in water suspension or other lipid-basedpharmaceutically acceptable carrier) are administered by oral gavage tomice with a volume of 5 mL/kg. The oral dosage of active compound is 500mg/kg mouse.

Before challenge, each group of mice receive oral dosages of activecompound or vehicle at 24 hours, 12 hours, and 2 hours before thechallenge.

After challenge, the same oral dosages of active compound or vehicle aregiven to each mouse 7 hours, 22 hours, 31 hours, 46 hours, and 55 hoursafter the challenge. The thickness of the left and right ears aremeasured before challenge, and 24, 48, and 72 hours after challenge.Results are expressed as net swelling: thickness after challenge minusthickness before challenge. Net swelling of treated mice vs. controlmice are compared.

Topical Administration:

Active compound 4-methylsulfonyl-2-butenenitrile prepared in vehicle(1:1; acetone:ethanol) at 375 mM and vehicle alone are topically appliedto both ears of the mice in a volume of 20 μl.

The topical doses are given after challenge to each mouse 7 hours, 22hours, 31 hours, 46 hours, and 55 hours after the challenge. Thethickness of the left and right ears are measured before challenge, and24, 48, and 72 hours after challenge. Results are expressed as netswelling: thickness after challenge minus thickness before challenge.Net swelling of treated mice vs. control mice are compared.

Example 13. Treatment of Atopic Dermatitis (Prophetic Example)

Objectives:

To investigate the efficacy of 4-methylsulfonyl-2-butenenitrile inpatients having atopic dermatitis.

Topical Formulation:

4-methylsulfonyl-2-butenenitrile is prepared as a gel formulationaccording to Example 2. Placebo contains the same gel ingredientswithout the active compound.

Oral Formulation:

Capsules or tablets each containing 100-800 mg of the active compound4-methylsulfonyl-2-butenenitrile are used in this example. Placebocapsules or tablets do not contain the active compound.

Methodology:

This is a randomized, double-blind, placebo controlled, paralleltreatment clinical activity study.

Male and female patients with mild to severe atopic dermatitis areenrolled after discontinuation of all treatments for atopic dermatitisfor a period of 4 weeks before study initiation. Patients are randomizedin a 1:1 ratio (active gel, placebo). A total of 300 patients areenrolled and treated.

The active gel or placebo is applied twice a day to affected areas ofthe body for 12 weeks.

The capsules or tablets are orally administered to patients 1-4 times aday for 12 weeks.

The treatment results are evaluated at 2 week intervals until week 12and then at 4 weeks after discontinuation of the study medication.

Criteria for Evaluation:

Safety:

Safety is evaluated by general history and physical signs, laboratorytesting for hematology, serum chemistry, and urinalysis, and byevaluations of local application site tolerability parameters oferythema, scaling, dryness, stinging/burning utilizing a rating scale of“0” (None) to “3” (Severe).

Efficacy:

Efficacy is Evaluated Utilizing:

1. an overall assessment of disease severity at study entry and at 2week intervals until week 12 and subsequently at 4 weeks after studymedication discontinuation. The investigator global assessment, IGA, isbased upon a rating scale of 0 to 4 with 0=none or clear, 1=almostclear, 2=mild disease involvement, 3=moderate disease involvement, and4=severe disease involvement, and:2. separate evaluation of a representative target atopic dermatitis areaof involvement for erythema, induration, lichenification, scaling, andoozing and crusting with each parameter rated on a 0-4 scale with 0=noneor clear, 1=almost clear, 2=mild disease involvement, 3=moderate diseaseinvolvement, and 4=severe disease involvement.

Statistical analyses of each of these efficacy evaluations are performedfor each of the 2 week study time points. Definitive evaluation ofefficacy is based upon comparisons of active to vehicle groups at end oftreatment at 12 weeks. The 4 week-post treatment evaluation is utilizedto evaluate durability of treatment effect after medicationdiscontinuation.

Example 14. Treatment of Psoriasis (Prophetic Example)

Objectives:

To investigate the efficacy of the 4-methylsulfonyl-2-butenenitrile inpatients having psoriasis vulgaris.

Topical Formulation:

4-methylsulfonyl-2-butenenitrile is prepared as a gel formulationaccording to Example 2.

Placebo contains the same gel ingredients without the active compound.

Oral Formulation:

Capsules or tablets each containing 100-800 mg of the active compound4-methylsulfonyl-2-butenenitrile are used in this example. Placebocapsules or tablets do not contain the active compound.

Methodology:

This is a randomized, double-blind, placebo controlled, paralleltreatment clinical activity study.

Male and female patients with mild to severe psoriasis vulgaris areenrolled. Patients discontinue all treatments for psoriasis for a periodof 4 weeks before study initiation. Patients are randomized in a 1:1ratio (active gel, placebo). A total of 200 patients are enrolled andtreated.

The active gel or placebo is applied twice a day to affected areas ofthe body for 12 weeks.

The capsules or tablets are orally administered to patients 1-4 times aday for 12 weeks.

The treatment results are evaluated at 2 week intervals until week 12and then at 4 weeks after discontinuation of the study medication.

Criteria for Evaluation:

Safety:

Safety is evaluated by general history and physical signs, laboratorytesting for hematology, serum chemistry, and urinalysis, and byevaluations of local application site tolerability parameters oferythema, scaling, dryness, stinging/burning utilizing a rating scale of“0” (None) to “3” (Severe).

Efficacy:

Efficacy is Evaluated Utilizing:

1. an overall assessment of disease severity at study entry and at 2week intervals until week 12 and subsequently at 4 weeks after studymedication discontinuation. The investigator global assessment, IGA, isbased upon a rating scale of 0 to 4 with 0=none or clear, 1=almostclear, 2=mild disease involvement, 3=moderate disease involvement, and4=severe disease involvement, and:2. separate evaluation of a representative target psoriasis lesion forerythema, scaling, and thickness of each parameter rated on a 0-4 scalewith 0=none or clear, 1=almost clear, 2=mild disease involvement,3=moderate disease involvement, and 4=severe disease involvement.

Statistical analyses of each of the efficacy evaluations are performedfor each of the 2 week study time points. Definitive evaluation ofefficacy is based upon comparisons of active to vehicle groups at end oftreatment at 12 weeks. The 4 week-post treatment evaluation is utilizedto evaluate durability of treatment effect after medicationdiscontinuation.

Example 15. Treatment of Acne (Prophetic Example)

Objectives:

To investigate the efficacy of the 4-methylsulfonyl-2-butenenitrile inpatients having acne vulgaris.

Topical Formulation:

4-methylsulfonyl-2-butenenitrile is prepared as a gel formulationaccording to Example 2. Placebo contains the same gel ingredientswithout the active compound.

Oral Formulation:

Capsules or tablets each containing 100-800 mg of the active compound4-methylsulfonyl-2-butenenitrile are used in this example. Placebocapsules or tablets do not contain the active compound.

Methodology:

This is a randomized, double-blind, placebo controlled, paralleltreatment clinical activity study.

Male and female patients with mild to severe acne vulgaris are enrolled.Patients discontinue all treatments for acne for a period of 4 weeksbefore initiation of the study. Patients are randomized in a 1:1 ratio(active gel, placebo). A total of 500 patients are enrolled and treated.

The active gel or placebo is applied twice a day to affected areas ofthe body for 12 weeks.

The capsules or tablets are orally administered to patients 1-4 times aday for 12 weeks.

The treatment results are evaluated at 2 week intervals until week 12and then at 4 weeks after discontinuation of the study medication.

Criteria for Evaluation:

Safety:

Safety is evaluated by general history and physical signs, laboratorytesting for hematology, serum chemistry, and urinalysis, and byevaluations of local application site tolerability parameters oferythema, scaling, dryness, stinging/burning utilizing a rating scale of“0” (None) to “3” (Severe).

Efficacy:

Efficacy is Evaluated Utilizing:

1. an overall assessment of disease severity at study entry and at 2week intervals until week 12 and subsequently at 4 weeks afterdiscontinuation of the study medication.The investigator global assessment, IGA, is based upon a rating scale of0 to 4 with 0=none or clear, 1=almost clear, 2=mild disease involvement,3=moderate disease involvement, and 4=severe disease involvement, and:2. separate counts of all types of acne lesions i.e. open and closedcomedones, papules, pustules, nodules, and cysts.

Statistical analyses of each of the efficacy evaluations are performedfor each of the 2 week study time points. Definitive evaluation ofefficacy is based upon comparisons of active to vehicle groups at end oftreatment at 12 weeks. The 4 week-post treatment evaluation is utilizedto evaluate durability of treatment effect after medicationdiscontinuation.

It is to be understood that the foregoing describes preferredembodiments of the present invention and that modifications may be madetherein without departing from the scope of the present invention as setforth in the claims.

What is claimed is:
 1. A method of treating inflammation or pain,comprising the steps of: administering to a subject suffering frominflammation or pain 4-methylsulfonyl-2-butenenitrile or apharmaceutically acceptable salt thereof, in an amount effective totreat inflammation or pain.
 2. The method according to claim 1, whereinsaid method reduces or alleviates the symptoms of localizedmanifestations of inflammation characterized by acute or chronicswelling, pain, or redness.
 3. The method according to claim 1, whereinsaid compound is administered by topical administration or oraladministration.
 4. The method according to claim 1, wherein saidinflammation and/or pain is associated with a skeletal or musculardisease or condition selected from the group consisting of:musculoskeletal sprains, musculoskeletal strains, tendonopathy,peripheral radiculopathy, osteoarthritis, degenerative joint disease,juvenile arthritis, gout, ankylosing spondylitis, psoriatic arthritis,system lupus erythematosus, costochondritis, tendonitis, bursitis,temporomandibular joint syndrome, and fibromyalgia.
 5. The methodaccording to claim 1, wherein said inflammation and/or pain isassociated with joints, ligaments, tendons, bone, muscles, or fascia. 6.The method according to claim 1, wherein said inflammation and/or painis associated with an inflammatory skin disease or disorder ofdermatitis or psoriasis.
 7. A method of treating an inflammatory skindisease or disorder, comprising the step of: administering to a subjectsuffering from an inflammatory skin disease or disorder4-methylsulfonyl-2-butenenitrile or a pharmaceutically acceptable saltthereof, in an amount effective to reduce or eliminate the symptoms ofthe inflammatory skin disease or disorder, wherein the inflammatory skindisease or disorder is dermatitis, psoriasis, acne, or rosacea.
 8. Themethod according to claim 7, wherein said method treats atopicdermatitis and alleviates one or more symptoms selected from the groupconsisting of erythema, induration, lichenification, scaling, and oozingand crusting.
 9. The method according to claim 7, wherein said methodtreats psoriasis and alleviates erythema, scaling, and/or thickness ofthe psoriasis lesions.
 10. The method according to claim 7, wherein saidmethod treats acne and alleviates one or more lesions selected from thegroups consisting of closed comedones, papules, pustules, nodules, andcysts.
 11. The method according to claim 7, wherein said method treatsrosacea and alleviates one or more symptoms selected from the groupconsisting of erythema, telangiectasia, red domed papules and pustules,red gritty eyes, and burning and stinging sensations.